Promoter sequences

Abstract

The present invention relates to an isolated human Cytochrome P-450 3A7 (CYP3A7) promoter region. The invention also relates to screening methods for agents modulating the expression of CYP3A7, such agents being potentially useful in modulating metabolism of endogenous and/or exogenous compounds, drug interaction, toxicity and/or bioavailability of drugs.

What is claimed is:  
     
         1 . An isolated human CYP3A7 promoter region comprising the dNR1 nucleotide sequence shown as positions 3425 to 3439 in SEQ ID NO: 1.  
     
     
         2 . An isolated human CYP3A7 promoter region comprising the dNR2 nucleotide sequence shown as positions 3469 to 3486 in SEQ ID NO: 1.  
     
     
         3 . The isolated human CYP3A7 promoter region of  claim 1 , comprising the XREM nucleotide sequence shown as positions 1832 to 3655 in SEQ ID NO: 1.  
     
     
         4 . An isolated human CYP3A7 promoter region comprising the dNR3 nucleotide sequence shown as positions 3855 to 3869 in SEQ ID NO: 1.  
     
     
         5 . The isolated human CYP3A7 promoter region of  claim 1 , comprising the nucleotide sequence set forth as SEQ ID NO: 1.  
     
     
         6 . A recombinant construct comprising the human CYP3A7 promoter region of  claim 1 .  
     
     
         7 . The recombinant construct of  claim 6 , wherein the human CYP3A7 promoter region is operably linked to a nucleic acid molecule comprising a nucleotide sequence that encodes a detectable product.  
     
     
         8 . The recombinant construct of  claim 7 , wherein the nucleic acid molecule comprises a sequence set forth as SEQ ID NO:2.  
     
     
         9 . A vector comprising the recombinant construct of  claim 6 .  
     
     
         10 . A host cell stably transformed with the vector of  claim 9 .  
     
     
         11 . A method for identification of an agent capable of modulating CYP3A7 promoter activity, the method comprising: 
 (i) contacting a candidate agent with the human CYP3A7 promoter region of  claim 1;  and    (ii) determining whether the candidate agent modulates CYP3A7 promoter activity.    
     
     
         12 . A method for identification of an agent capable of modulating CYP3A7 promoter activity, the method comprising: 
 (i) providing a cell comprising the human CYP3A7 promoter region of  claim 1 , operably linked to a reporter gene;    (ii) determining a first level of expression of the reporter gene;    (iii) contacting the cell with a candidate agent; and    (iv) determining a second level of expression of the reporter gene in the presence of the candidate agent, wherein a difference between the first and second levels of expression indicates that the candidate agent modulates CYP3A7 promoter activity.    
     
     
         13 . A method for identifying an agent that modulates the association of a CYP3A7 promoter and a transcription factor, the method comprising: 
 (i) contacting the CYP3A7 promoter region of  claim 1  with a transcription factor;    (ii) determining a first association between the CYP3A7 promoter region and the transcription factor;    (iii) contacting the CYP3A7 promoter region and the transcription factor with a candidate agent; and    (iv) determining a second association between the CYP3A7 promoter region and the transcription factor in the presence of the candidate agent, wherein a difference between the first and the second association indicates that the candidate agent modulates the association of the CYP3A7 promoter and the transcription factor.    
     
     
         14 . The method of  claim 13 , wherein the method is a solid phase in vitro binding assay.  
     
     
         15 . The method of  claim 13 , wherein the method is a cell based transcription assay.  
     
     
         16 . A method for identifying an agent that modulates the association of a CYP3A7 promoter and a transcription factor, the method comprising: 
 (i) providing a cell comprising the CYP3A7 promoter region of  claim 1 , operably linked to a reporter gene, and the transcription factor;    (ii) determining a first level of expression of the reporter gene;    (iii) contacting the cell with a candidate agent; and    (iv) determining a second level of expression of the reporter gene in the presence of the candidate agent, wherein a difference between the first and second levels of expression indicates that the candidate agent modulates the association of the CYP3A7 promoter and the transcription factor.    
     
     
         17 . The method according to  claim 13 , wherein the transcription factor is the Pregnane Activated Receptor (PAR).  
     
     
         18 . The method according to  claim 13 , wherein the transcription factor is the Constitutive Androstane Receptor (CAR).  
     
     
         19 . The isolated human CYP3A7 promoter region of  claim 1 , comprising the XREM nucleotide sequence shown as positions 3330 to 3551 in SEQ ID NO:1.  
     
     
         20 . The isolated human CYP3A7 promoter region of  claim 1 , further comprising the dNR2 nucleotide sequence shown as positions 3469 to 3486 in SEQ ID NO: 1.  
     
     
         21 . The isolated human CYP3A7 promoter region of  claim 20 , further comprising the dNR3 nucleotide sequence shown as positions 3855 to 3869 in SEQ ID NO: 1.  
     
     
         22 . A nucleic acid comprising a nucleotide sequence selected from the group consisting of nucleotides 3425 to 3439 of SEQ ID NO:1, nucleotides 3469 to 3486 of SEQ ID NO:1, nucleotides 3855 to 3869 of SEQ ID NO:1, and nucleotides 3330 to 3551 of SEQ ID NO: 1, operably linked to a heterologous coding sequence.